Substrate access path-guided engineering of <scp>l</scp>-threonine aldolase for improving diastereoselectivity

Abstract

The L-threonine aldolase from <i>Leishmania major</i> was engineered to improve its diastereoselectivity by a CAST/ISM strategy, providing insights into the relationship between the physico-chemical properties of the substrate access path and diastereoselectivity. The steric hindrance, hydrophobic interaction and π-π interaction cooperated to improve the diastereoselectivity of the enzyme, with a diastereomeric excess (de) value reaching 96.3%_<i>syn</i> from 26.8%_<i>syn</i>.